An applied printing immunoassay with recombinant Nc-SAG1 for detection of antibodies to Neospora caninum in cattle

Abstract

Abstract. Neospora caninum is a protozoan parasite that causes an important reproductive disease in cattle. Neospora caninum surface antigen 1 (Nc-SAG1) is an immunodominant candidate for the development of a diagnostic reagent for neosporosis. The current study describes the development and evaluation of an antigen print immunoassay (APIA) with recombinant Nc-SAG1 for the detection of specific antibodies to N. caninum in cattle. The concordance between APIA and a commercial enzyme-linked immunosorbent assay (ELISA) was evaluated with 232 serum samples from experimentally and naturally infected cattle. Sixty-one (26.7%) samples were positive for antibodies to N. caninum by ELISA and 58 (25.4%) by APIA. The new assay had a sensitivity of 85% and a specificity of 96%. These results, along with the potential of APIA to evolve into a multiple antigen detection format, suggest that this method would be a reliable diagnostic test for detection of antibodies to N. caninum in cattle.

Cell mediated immune responses in the placenta following challenge of vaccinated pregnant heifers with Neospora caninum

“…The NC-1 strain of N. caninum was used for the cloning and purification of the recombinant proteins NcSAG1 (rNcSAG1), NcHSP20 (rNcHSP20) and NcGRA7 (rNcGRA7) (Wilkowsky et al, 2011) that were used to immunize group C heifers, as described by Hecker et al (2014). All heifers were iv-challenged with 4.7 × 10 7 live NC-1 tachyzoites in a final volume of 3 ml of PBS (Hecker et al, 2013(Hecker et al, , 2014.…”

Validation and field evaluation of a competitive inhibition ELISA based on the recombinant protein tSAG1 to detect anti-Neospora caninum antibodies in sheep and goats

“…An applied printing immunoassay (APIA) and an iELISA based on recombinant tSAG1 protein were previously evaluated for use in cattle (Chahan et al, 2003;Wilkowsky et al, 2011). A Se of 85% and a Sp of 96% relative to the commercial iELISA IDEXX Neospora Ab were reported for APIA (Wilkowsky et al, 2011). The iELISA reported a Se of 100 % and a Sp of 95,8% relative to IB (Chahan et al, 2003).…”

Neosporosis bovina en Argentina: a 25 años del primer reporte en el país

“…A continuación, se resumen los principales aportes realizados. Wilkowsky et al (2011) desarrollaron una prueba inmunoenzimática en línea (applied printing immunoassay -APIA-) para la detección de anticuerpos anti-N. caninum en bovinos, con la proteína recombinante Nc-SAG1 como antígeno. Los autores propusieron su uso como prueba de screening, dada su especificidad diagnóstica (DSp) y facilidad en su lectura cualitativa dicotómica, que podría ser aplicable a estudios de prevalencia.…”